Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan 250012, China; Department of Obstetrics and Gynecology, Hangzhou Women's Hospital & Hangzhou Maternity and Child Health Care Hospital, Hangzhou 310000, China.
Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan 250012, China.
Gene. 2018 Aug 30;669:99-106. doi: 10.1016/j.gene.2018.05.087. Epub 2018 May 23.
Increasing evidence demonstrates that the four and a half LIM domain (FHL) gene and its protein products have different functions in the progression of various malignancies. However, the role of FHL protein 2 (FHL2) in cervical cancer (CC) has not been fully elucidated. In this study, we investigated the prognostic value of FHL2 expression in human CC tissues and the potential molecular mechanisms through which FHL2 modulates CC cell proliferation and apoptosis.
We measured FHL2 expression in CC cell lines and tissues by quantitative real-time polymerase chain reaction and Western blot assays. The effects of FHL2 knockdown on cell proliferation and apoptosis in two CC cell lines were examined using RNA interference, cell counting kit-8, Western blot and flow cytometry assays. Furthermore, we assessed phosphorylated protein kinase B (p-AKT) and phosphorylated mammalian target of rapamycin (p-mTOR) expression in two CC cell lines to determine whether the AKT/mTOR pathway is involved in the effects of FHL2 silencing on cell proliferation and apoptosis. Nude mice tumorigenicity experiments were also performed to evaluate the effects of FHL2 on HeLa cell growth in vivo.
We found that FHL2 was significantly upregulated in CC cell lines and tissues. According to survival curves, high FHL2 expression levels in patients were correlated with poor prognosis. Moreover, by decreasing p-AKT and p-mTOR protein levels, silencing FHL2 significantly inhibited cell proliferation and induced apoptosis. FHL2 knockdown also induced apoptosis by increasing the Bax-to-Bcl2 ratio. By contrast, FHL2 overexpression significantly promoted cell proliferation. Finally, decreased tumour growth in an in vivo animal model also demonstrated the tumour-suppressing effects of FHL2 knockdown.
Our findings indicate that FHL2 is an important prognostic factor in CC and that it plays a crucial oncoprotein role by promoting cell proliferation and inhibiting apoptosis in CC, possibly by targeting the AKT/mTOR pathway.
越来越多的证据表明，四个半 LIM 结构域（FHL）基因及其蛋白产物在各种恶性肿瘤的进展中具有不同的功能。然而，FHL 蛋白 2（FHL2）在宫颈癌（CC）中的作用尚未完全阐明。在本研究中，我们调查了 FHL2 表达在人 CC 组织中的预后价值，以及 FHL2 调节 CC 细胞增殖和凋亡的潜在分子机制。
我们通过定量实时聚合酶链反应和 Western blot 检测 CC 细胞系和组织中的 FHL2 表达。使用 RNA 干扰、细胞计数试剂盒-8、Western blot 和流式细胞术检测 FHL2 敲低对两种 CC 细胞系中细胞增殖和凋亡的影响。此外，我们评估了两种 CC 细胞系中磷酸化蛋白激酶 B（p-AKT）和磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）的表达，以确定 AKT/mTOR 通路是否参与 FHL2 沉默对细胞增殖和凋亡的影响。还进行了裸鼠肿瘤发生实验，以评估 FHL2 对 HeLa 细胞体内生长的影响。
我们发现 FHL2 在 CC 细胞系和组织中显著上调。根据生存曲线，患者中 FHL2 表达水平较高与预后不良相关。此外，通过降低 p-AKT 和 p-mTOR 蛋白水平，沉默 FHL2 显著抑制细胞增殖并诱导细胞凋亡。FHL2 敲低还通过增加 Bax-to-Bcl2 比值诱导细胞凋亡。相比之下，FHL2 过表达显著促进细胞增殖。最后，体内动物模型中肿瘤生长的减少也证明了 FHL2 敲低的肿瘤抑制作用。
我们的研究结果表明，FHL2 是 CC 的一个重要预后因素，它通过促进 CC 中的细胞增殖和抑制凋亡发挥关键癌蛋白作用，可能通过靶向 AKT/mTOR 通路。
